DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Claim 76 was inadvertently placed in Group IV, but depends from claim 26 and so for clarity of the record is considered part of Group II for the purposes of restriction and Applicant’s election.
Applicant's election with traverse of Group II, presently claims 22, 26-29, 33, 39, and 76, the reply filed on 9/03/2025 is acknowledged. The traversal is on the ground(s) that the instant amendments to claims 1 and 22 restore unity of invention relative to the cited teachings of Saridaki. This is not found persuasive for several reasons. First, Group III still lacks unity of invention with Groups I, II, and IV because Group III does not share the same or corresponding technical feature. Groups, I, II, and IV lack unity of invention even though the inventions of these groups require a composition comprising keratinocytes deficient in the expression of TP53, CDKN2A, and NOTCH1, this technical feature is not a special technical feature as it does not make a contribution over the prior art in view of Su et al. (Theranostics (2017), 7(5), 1088-1099; Reference U) as evidenced by Walters and Roberts (Chapter 1 in Dermatological and Transdermal Formulations (2002), Taylor and Francis Group, ebook ISBN: 9780429164323; Reference V).
Claim 22 is a product-by-process claim. The patentability of product-by-process claims depends only on the structure of the claimed composition or any structural differences imparted to the final product by method steps in a product-by-process claim. See M.P.E.P. § 2113. In the instant case and absent evidence to the contrary, a prima facie case exists that the composition claimed in the product-by-process claims is substantially or completely identical to composition made by other methods. For the purposes of restriction, claim 22 is read as a composition of matter claim for the statutory class of invention. See M.P.E.P. § 2106.
Su teaches a human organisms suffering from oral squamous cell carcinoma and carrying mutations in genomic DNA of the TP53, CDKN2A, and NOTCH1 genes (Abstract; see p1089-1090 for DNA sample collection and sequencing; see the paragraph spanning p1091-1092). Su teaches that the TP53 mutations are loss-of-function (see the paragraph spanning both columns on p1097 as Su teaches restoring p53 function). Su teaches that the CDKN2A mutations are copy loss mutations (the paragraph spanning both columns on p1094). Su teaches that the majority of NOTCH1 mutations were inactivating (the paragraph spanning both columns on p1095).
Regarding claim 22, Walters and Roberts teach that animals inherently comprise keratinocytes (see the 1st paragraph of subheading III on the 14th page). In so much that Su teaches that the gene mutations for TP53, CDKN2A, and NOTCH 1 are in genomic DNA, these mutations are reasonably presumed present in all of the somatic cells of the human organisms of Su and inclusive of the keratinocytes in said human organisms. See M.P.E.P. § 2112.
Therefore, the keratinocytes of Su as evidenced by Walters and Roberts breaks unity of invention a posteriori and restriction is still necessitated between Groups I-IV.
The requirement is still deemed proper and is therefore made FINAL.
Claims 1, 3, 5, 6, 8-10, 15-17, 62, 65, and 75 are withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to nonelected inventions, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 9/03/2025.
Claims 22, 26-29, 33, 39, and 76 are under consideration on the merits.
Nucleotide and/or Amino Acid Sequence Disclosures
REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiencies and the required response to this Office Action are as follows:
Specific deficiency - This application fails to comply with the requirements of 37 CFR 1.821 - 1.825 because it does not contain a "Sequence Listing" as a separate part of the disclosure or a CRF of the “Sequence Listing.”
Required response - Applicant must provide:
A "Sequence Listing" part of the disclosure; together with
An amendment specifically directing its entry into the application in accordance with 37 CFR 1.825(a)(2);
A statement that the "Sequence Listing" includes no new matter as required by 37 CFR 1.821(a)(4); and
A statement that indicates support for the amendment in the application, as filed, as required by 37 CFR 1.825(a)(3).
If the "Sequence Listing" part of the disclosure is submitted according to item 1) a) or b) above, Applicant must also provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required incorporation-by-reference paragraph, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
If the "Sequence Listing" part of the disclosure is submitted according to item 1) c) or d) above, applicant must also provide:
A CRF in accordance with 37 CFR 1.821(e)(1) or 1.821(e)(2) as required by 1.825(a)(5); and
A statement according to item 2) a) or b) above.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 33 and 39 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 33, which depends from independent claim 22, recites “further comprising” but then adds a device-type limitation to the claim. Thus, it is unclear what composition the claim is referring to since the last line is referring only to the cells of claim 22 and not the composition per se. Furthermore, as phrased it is unclear if claim 33 is claiming a device, a composition, or both; the preamble of claim 22 is directed towards a composition but claim 33 adds device-type limitations. Clarification and/or correction is required.
Claim 39 recites product-by-process limitations towards freezing and which depends from composition claim 33, and is indefinite because it is unclear if the claim is limiting the scope to the process of freezing or the frozen product itself. Clarification and/or correction is required.
In so much that claim 39 depends from claim 33 and does not resolve the point of confusion, this claim must also be rejected with claim 33 as indefinite for the reasons given above.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
Claims 22, 26-29, 33, 39, and 76 are rejected under 35 U.S.C. 101 because the claimed invention is directed to natural product without significantly more.
Independent claim 22 recites a natural product, keratinocytes that are deficient in expression of TP53, CDKN2A, and NOTCH1. Su (Theranostics (2017), 7(5), 1088-1099; Reference U) teaches a human organisms suffering from oral squamous cell carcinoma and carrying mutations in genomic DNA of the TP53, CDKN2A, and NOTCH 1 genes (Abstract; see p1089-1090 for DNA sample collection and sequencing; see the paragraph spanning p1091-1092). Su teaches that the TP53 mutations are loss-of-function (see the paragraph spanning both columns on p1097 as Su teaches restoring p53 function). Su teaches that the CDKN2A mutations are copy loss mutations (the paragraph spanning both columns on p1094). Su teaches that the majority of NOTCH1 mutations were inactivating (the paragraph spanning both columns on p1095). Walters and Roberts (Chapter 1 in Dermatological and Transdermal Formulations (2002), Taylor and Francis Group, ebook ISBN: 9780429164323; Reference V) teach that animals inherently comprise keratinocytes (see the 1st paragraph of subheading III on the 14th page). In so much that Su teaches that the gene mutations for TP53, CDKN2A, and NOTCH 1 are in genomic DNA, these mutations are reasonably presumed present in all of the somatic cells of the human organisms of Su and inclusive of the keratinocytes in said human organisms. See M.P.E.P. § 2112.
“genome-edited” in claim 22 is a product-by-process limitation and no specific mutations are claimed that might otherwise be markedly different from the naturally occurring mutations in keratinocytes of Su as evidenced by Walters and Roberts. Therefore, the composition of claim 22 does not appear to possess any markedly different characteristic as compared to the nearest natural counterpart. See M.P.E.P. § 2106.04(c)(I).
In so much that Su teaches that the keratinocytes that are deficient in expression of TP53, CDKN2A, and NOTCH1 are present in human organisms, the broadest reasonable interpretation of claim 22 encompasses human organisms and so the claim is also not patent eligible as humans per se, which are excluded under The Leahy-Smith America Invents Act (AIA ), Public Law 112-29, sec. 33, 125 Stat. 284 (September 16, 2011). See M.P.E.P. § 2106.03(I).
Claims 26, 27, and 76 depend from claim 22. Walters and Roberts teach that the skin of animals inherently comprises a top layer of keratinocytes in the stratum corneum (see the 1st paragraph of subheading III on the 14th page) and a bottom layer of tissue comprising the dermis wherein the dermis comprises extracellular matrix (e.g. the basement membrane comprising type IV collagen, laminin, nidogen, and fibronectin) and fibroblasts (Fig. 3 and 4; the paragraph spanning the 6th-7th page; the 1st paragraph of subheading “B. dermis” on the 11th page). Therefore, claims 26, 27, and 76 are directed towards the natural product of the keratinocytes of Su carrying mutations in TP53, CDKN2A, and NOTCH1 as evidenced by Walters and Roberts and which do not appear to possess any markedly different characteristic as compared to the nearest natural counterpart. See M.P.E.P. § 2106.04(c)(I).
This judicial exception is not integrated into a practical application because claims 22, 26, 27, and 76 are only directed towards the natural product and the claimed natural product does not appear to possess any markedly different characteristic as compared to the nearest natural counterpart.
Regarding claims 28, 29, 33, and 39, the claims do not include additional elements that are sufficient to amount to significantly more than the judicial exception because the additional elements are routine and conventional in this art.
Regarding claims 28 and 29, it would be routine and conventional in this art to further add a Matrigel layer as a species of hydrogel which inherently comprises growth factors in view of Scott et al. (Journal of Visualized Experiments (2011), 58, e3525, 5 pages; Reference V2) and as evidenced by Vukicevic et al. (Experimental Cell Research (1992), 202, 1-8; Reference W2). Vukicevic teaches that Matrigel inherently comprises a plurality of growth factors (Abstract). Scott teaches an inverse Matrigel invasion assay, wherein Matrigel is deposited on both sides of a Transwell filter insert and a chemoattractant is deposited on top of the Transwell filter insert to induce cellular invasion/migration (p2, subheading 2), reading on claims 28 and 33. Scott teaches the inverse Matrigel invasion assay is advantageous to generate considerably more information about cellular invasion, such as being able to visualize differently labeled cells to examine leading versus following cells in collective invasion using this method allows cells to be fixed, permeabilized, stained and then imaged for protein levels or subcellular distribution (Figure 3C and 3D and Discussion on p5), reading on claim 33.
Regarding claim 33, it would be routine and conventional in this art to further add the embodiment of a Transwell filter insert nested within a cell culture well plate in view of Wojtowicz (Wound Rep Reg (2014) 22 246–255; Reference X). Wojtowicz teaches a bilayered living cellular construct (BLCC; Abstract), which is made in a coculture system comprising human keratinocytes and human dermal fibroblasts, wherein the fibroblasts (e.g. tissue) are mixed with type I collagen are first deposited on the top surface of the porous membrane of a Transwell filter (and wherein the membrane defines a planar surface that is present in both the top and bottom chambers) and then the keratinocytes are deposited on top of the fibroblast layer (Fig. 1 and the paragraph spanning p247-248), and reading in-part on the single insert device having an upper and bottom chamber in fluid communication and being separated by a ledge and the embodiment of collagen as a biodegradable polymer for claim 33. Further addition of Matrigel as a species of hydrogel positioned in the bottom chamber would be routine and conventional over Scott for the reasons given above.
Regarding claim 39, it would be routine and conventional in this art to further freeze keratinocytes in view of Johansen et al (Journal of Visualized Experiments (2017), 130, e56863, 5 pages; Reference X2) as evidenced by Haynes et al. (CRC Handbook of Chemistry and Physics, 97th Edition (2017), ISBN 978-1-4987-5429-3; Section 4; Reference U3). Johansen teaches freezing and then storing primary keratinocytes in liquid nitrogen until ready for use (p2, subheading 3), and wherein nitrogen is a liquid at about -195.8 to about -252 °C as evidenced by Haynes (see “Nitrogen” in Section 4, 4-24), reading on claim 39.
For the reasons given above, claims 22, 26-29, 33, 39, and 76 are rejected as patent ineligible under 35 U.S.C. § 101.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 22, 26, 27, and 76 are rejected under 35 U.S.C. 102(a)(1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Su et al. (Theranostics (2017), 7(5), 1088-1099; Reference U) as evidenced by Walters and Roberts (Chapter 1 in Dermatological and Transdermal Formulations (2002), Taylor and Francis Group, ebook ISBN: 9780429164323; Reference V).
Su teaches a human organisms suffering from oral squamous cell carcinoma and carrying mutations in genomic DNA of the TP53, CDKN2A, and NOTCH 1 genes (Abstract; see p1089-1090 for DNA sample collection and sequencing; see the paragraph spanning p1091-1092). Su teaches that the TP53 mutations are loss-of-function (see the paragraph spanning both columns on p1097 as Su teaches restoring p53 function). Su teaches that the CDKN2A mutations are copy loss mutations (the paragraph spanning both columns on p1094). Su teaches that the majority of NOTCH1 mutations were inactivating (the paragraph spanning both columns on p1095).
Regarding claim 22, Walters and Roberts teach that animals inherently comprise keratinocytes (see the 1st paragraph of subheading III on the 14th page). In so much that Su teaches that the gene mutations for TP53, CDKN2A, and NOTCH 1 are in genomic DNA, these mutations are reasonably presumed present in all of the somatic cells of the human organisms of Su and inclusive of the keratinocytes in said human organisms. Therefore, the human organisms of Su as evidenced by Walters and Roberts anticipates or reads on the composition of claim 22.
Regarding claims 26, 27, and 76, Walters and Roberts teaches that the skin of animals inherently comprises a top layer of keratinocytes in the stratum corneum (see the 1st paragraph of subheading III on the 14th page) and a bottom layer of tissue comprising the dermis wherein the dermis comprises extracellular matrix (e.g. the basement membrane comprising type IV collagen, laminin, nidogen, and fibronectin) and fibroblasts (Fig. 3 and 4; the paragraph spanning the 6th-7th page; the 1st paragraph of subheading “B. dermis” on the 11th page). Therefore, the human organisms of Su as evidenced by Walters and Roberts anticipates or reads on the composition of claims 26, 27, and 76.
Claim 22 is a product-by-process claims, reciting limitations to “genome-edited” keratinocyte. See M.P.E.P. § 2113; product-by-process claims are not limited to the manipulations of the recited steps, only the structure implied by the steps. Furthermore, alternate grounds of rejection under both 102 and 103 is permissible given the lack of physical description of product-by-process claims and the inability of the USPTO to manufacture and compare products. See M.P.E.P. § 2113 (III). Once a product appearing to be substantially identical is found and an art rejection made, the burden shifts to the applicant to show an unobvious difference. In this case, the burden is shifted to Applicant to show that the manufacturing process steps of the product-by-process claims impart any novel and/or non-obvious structural characteristics to the claimed product as compared to the composition taught by Su as evidenced by Walters and Roberts. Particularly, if the product-by process limitations of claim 22 impart no structural difference then the claim is anticipated. If the product-by process limitations of claim 22 impart a structural difference, then Applicant must clearly set forth why any structural difference between the claimed composition and the composition of Su as evidenced by Walters and Roberts is non-obvious.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claims 22, 26, 27, and 76 are rejected under 35 U.S.C. 103 as being unpatentable over Indra et al. (Hormone Research (2000), 54, 296-300; provided in the IDS dated 11/03/2022) in view of Cong et al. (US 9,370,551; Reference A) and Gurudutt et al. (Journal of Skin Cancer (2011), article ID 502723, 10 pages; Reference W) and as evidenced by Walters and Roberts (Chapter 1 in Dermatological and Transdermal Formulations (2002), Taylor and Francis Group, ebook ISBN: 9780429164323; Reference V).
Indra teaches transgenic mice comprising somatic mutations in epidermal keratinocytes (Abstract; see the Materials and Methods of 1st paragraph of the Results and Discussion on p297-298), reading in-part on claim 22.
Regarding claim 22, Walters and Roberts teach that animals inherently comprise keratinocytes (see the 1st paragraph of subheading III on the 14th page). In so much that Su teaches that the gene mutations for TP53, CDKN2A, and NOTCH 1 are in genomic DNA, these mutations are reasonably presumed present in all of the somatic cells of the human organisms of Su and inclusive of the keratinocytes in said human organisms. Therefore, the mice of Indra as evidenced by Walters and Roberts reads in-part on the composition of claim 22. Indra teaches that the transgenic mice would be useful as an animal model of skin diseases (the Conclusion on p300), reading in-part on claim 22.
Regarding claims 26, 27, and 76, Walters and Roberts teaches that the skin of animals inherently comprises a top layer of keratinocytes in the stratum corneum (see the 1st paragraph of subheading III on the 14th page) and a bottom layer of tissue comprising the dermis wherein the dermis comprises extracellular matrix (e.g. the basement membrane comprising type IV collagen, laminin, nidogen, and fibronectin) and fibroblasts (Fig. 3 and 4; the paragraph spanning the 6th-7th page; the 1st paragraph of subheading “B. dermis” on the 11th page). Therefore, the mice of Indra as evidenced by Walters and Roberts reads in-part on the composition of claims 26, 27, and 76.
Regarding claim 22, Indra does not teach keratinocytes deficient in expression of TP53, CDKN2A, and NOTCH1.
Cong teaches methods of treating head and neck squamous cell carcinoma (HNSCC) by altering the expression of HNSCC-related genes and gene expression (Abstract). Cong teaches that knockdown or deregulated expression of NOTCH1 leads to a differentiation block and increased proliferation of keratinocytes (Col 12, lines 27-41) and that point mutations in the NOTCH1 gene occur in 11% of HNSCC tumors (Col. 11, lines 39-58), reading on claim 22. Cong teaches that TP53 is the most commonly mutated gene in HNSCC and teaches a 100 kb deletion of TP53 (Col. 11, lines 16-38), reading on claim 22. Cong teaches CDKN2A deletions (Col. 10, lines 14-28), reading on claim 22. Cong teaches genes with aberrant loss-of-function or gain-of-function comprising NOTCH1, CDKN2A, and TP53 (Col. 2, line 59 through Col. 3, line 9and Col. 3, lines 61-65), reading on claim 22. Cong teaches altering the expression of NOTCH1, CDKN2A, or TP53 in a mammalian cell (claim 1), reading on claim 22.
Gurudutt teaches that cutaneous squamous cell carcinoma (of the head and neck) a type of non-melanoma skin cancer and is the second most common skin malignancy in the US (Abstract and Introduction), reading on claim 22.
Regarding claim 22, It would have been obvious to a person of ordinary skill in the art before the invention was filed to add the known mutations deficient in expression of TP53, CDKN2A, and NOTCH1 taught by Cong to the keratinocytes of Indra in view of Gurudutt. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because Indra teaches detailed methods for introducing somatic mutations only to the keratinocytes of mice and envisions utilizing the transgenic mice as a model for skin disease, Cong teaches known mutations in TP53, CDKN2A, and NOTCH1 in head and neck squamous cell carcinoma and keratinocyte function is altered when NOTCH1 is knocked down or has deregulated gene expression, and Gurudutt teaches that head and neck squamous cell carcinoma is also a type of non-melanoma skin cancer. The skilled artisan would have been motivated to do so because introducing the mutations deficient in expression of TP53, CDKN2A, and NOTCH1 taught by Cong would predictably improve the keratinocytes and transgenic mice of Indra as a model for skin disease and particularly cutaneous squamous cell carcinoma.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claims 22, 26-29, 33, and 76 are rejected under 35 U.S.C. 103 as being unpatentable over Wojtowicz et al. (Wound Rep Reg (2014) 22 246–255; Reference X) in view of Cong et al. (US 9,370,551; Reference A), Gurudutt et al. (Journal of Skin Cancer (2011), article ID 502723, 10 pages; Reference W), Bajaj et al. (Molecular Therapy (2005), 11(6), 969-979; Reference U2), and Scott et al. (Journal of Visualized Experiments (2011), 58, e3525, 5 pages; Reference V2) and as evidenced by Vukicevic et al. (Experimental Cell Research (1992), 202, 1-8; Reference W2)
In view of the 35 U.S.C. § 112(d) rejection above and in the interest of compact prosecution, claim 33 is treated as depending from claim 22.
Wojtowicz teaches a bilayered living cellular construct (BLCC; Abstract), which is made in a coculture system comprising human keratinocytes and human dermal fibroblasts, wherein the fibroblasts (e.g. tissue) are mixed with type I collagen are first deposited on the top surface of the porous membrane of a Transwell filter (and wherein the membrane defines a planar surface that is present in both the top and bottom chambers) and then the keratinocytes are deposited on top of the fibroblast layer (Fig. 1 and the paragraph spanning p247-248), reading in-part on claim 22, reading on claim 26 and the embodiment of collagen for an extracellular protein, claim 27, reading in-part on claims 28 and 29, and reading in-part on the single insert device having an upper and bottom chamber in fluid communication and being separated by a ledge and the embodiment of collagen as a biodegradable polymer for claim 33, and reading on claim 76.
Regarding claim 22, Wojtowicz does not teach genome-edited keratinocytes deficient in expression of TP53, CDKN2A, and NOTCH1. Regarding claim 28, Wojtowicz does not teach the embodiment of Matrigel as a species of hydrogel. Regarding claim 29, Wojtowicz does not teach a hydrogel further comprising a plurality of growth factors. Regarding claim 33, Wojtowicz does not teach the embodiment of Matrigel as a species of hydrogel positioned in the bottom chamber.
Cong teaches methods of treating head and neck squamous cell carcinoma (HNSCC) by altering the expression of HNSCC-related genes and gene expression (Abstract). Cong teaches that knockdown or deregulated expression of NOTCH1 leads to a differentiation block and increased proliferation of keratinocytes (Col 12, lines 27-41) and that point mutations in the NOTCH1 gene occur in 11% of HNSCC tumors (Col. 11, lines 39-58), reading on claim 22. Cong teaches that TP53 is the most commonly mutated gene in HNSCC and teaches a 100 kb deletion of TP53 (Col. 11, lines 16-38), reading on claim 22. Cong teaches CDKN2A deletions (Col. 10, lines 14-28), reading on claim 22. Cong teaches genes with aberrant loss-of-function or gain-of-function comprising NOTCH1, CDKN2A, and TP53 (Col. 2, line 59 through Col. 3, line 9 and Col. 3, lines 61-65), reading on claim 22. Cong teaches altering the expression of NOTCH1, CDKN2A, or TP53 in a mammalian cell (claim 1), reading on claim 22.
Gurudutt teaches that cutaneous squamous cell carcinoma (of the head and neck) a type of non-melanoma skin cancer and is the second most common skin malignancy in the US (Abstract and Introduction), reading on claim 22.
Bajaj teaches detailed methods of generating genetically-modified keratinocytes by retrovirus transduction (p977, subheadings “Retrovirus production and cell culture” and “Transduction methods”), reading in-part on the genome-edited keratinocytes of claim 22.
Scott teaches an inverse Matrigel invasion assay, wherein Matrigel is deposited on both sides of a Transwell filter insert and a chemoattractant is deposited on top of the Transwell filter insert to induce cellular invasion/migration (p2, subheading 2), reading on claims 28 and 33. Scott teaches the inverse Matrigel invasion assay is advantageous to generate considerably more information about cellular invasion, such as being able to visualize differently labeled cells to examine leading versus following cells in collective invasion using this method allows cells to be fixed, permeabilized, stained and then imaged for protein levels or subcellular distribution (Figure 3C and 3D and Discussion on p5), reading on claim 33.
Vukicevic teaches that Matrigel inherently comprises a plurality of growth factors (Abstract). Therefore, Scott as evidenced by Vukicevic reads on claim 29.
Regarding claim 22, It would have been obvious to a person of ordinary skill in the art before the invention was filed to add the known mutations deficient in expression of TP53, CDKN2A, and NOTCH1 taught by Cong to the keratinocytes of Wojtowicz in view of Gurudutt and Bajaj. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because Cong teaches known mutations in TP53, CDKN2A, and NOTCH1 in head and neck squamous cell carcinoma and keratinocyte function is altered when NOTCH1 is knocked down or has deregulated gene expression, Gurudutt teaches that head and neck squamous cell carcinoma is also a type of non-melanoma skin cancer, and Bajaj teaches detailed methods of generating genetically-modified keratinocytes by retrovirus transduction. The skilled artisan would have been motivated to do so because introducing the mutations deficient in expression of TP53, CDKN2A, and NOTCH1 taught by Cong would predictably improve the keratinocytes of Wojtowicz as a model for skin disease and particularly cutaneous squamous cell carcinoma.
Regarding claims 28, 29, and 33, it would have been obvious to a person of ordinary skill in the art before the invention was filed to substitute the collagen of Wojtowicz with the Matrigel of Scott as evidenced by Vukicevic and further add the Matrigel of Scott to the underside of the Transwell filter inserts of Wojtowicz. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because both Wojtowicz and Scott are directed towards cellular coculture systems comprising Transwell filter inserts. The skilled artisan would have been motivated to do so because Scott teaches the inverse Matrigel invasion assay is advantageous to generate considerably more information about cellular invasion, such as being able to visualize differently labeled cells to examine leading versus following cells in collective invasion using this method allows cells to be fixed, permeabilized, stained and then imaged for protein levels or subcellular distribution and because Cong teaches at least some HNSCC mutations affect the functional properties of keratinocytes in vitro, the substitution and addition of Matrigel would be predictably advantageous to investigate the effects of the HNSCC mutations of Cong on the functional properties of the keratinocytes of Wojtowicz in the in vitro keratinocyte and fibroblast coculture system of Wojtowicz.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Claim 39 is rejected under 35 U.S.C. 103 as being unpatentable over Wojtowicz, Cong, Gurudutt, Bajaj, and Scott as evidenced by Vukicevic as applied to claims 22 and 33 above, and further in view of Johansen et al (Journal of Visualized Experiments (2017), 130, e56863, 5 pages; Reference X2) as evidenced by Haynes et al. (CRC Handbook of Chemistry and Physics, 97th Edition (2017), ISBN 978-1-4987-5429-3; Section 4; Reference U3).
The teachings of Wojtowicz, Cong, Gurudutt, Bajaj, and Scott as evidenced by Vukicevic are relied upon as set forth above.
Regarding claim 39, in view of the indefiniteness rejections above and in the interest of compact prosecution, Wojtowicz, Cong, Gurudutt, Bajaj, and Scott as evidenced by Vukicevic do not teach the embodiment of the keratinocyte composition of claims 22 and 33 further frozen between -20 to -212 degrees Celsius.
Johansen teaches freezing and then storing primary keratinocytes in liquid nitrogen until ready for use (p2, subheading 3), and wherein nitrogen is a liquid at about -195.8 to about -252 °C as evidenced by Haynes (see “Nitrogen” in Section 4, 4-24), reading on claim 39.
It would have been obvious to a person of ordinary skill in the art before the invention was filed to further freeze the keratinocyte and fibroblast composition of Wojtowicz in view of Johansen and as evidenced by Haynes. A person of ordinary skill in the art would have had a reasonable expectation of success to do so because both Wojtowicz and Johansen are in-part directed towards keratinocyte compositions. The skilled artisan would have been motivated to do so because Johansen teaches that freezing keratinocytes in liquid nitrogen is predicably advantageous to store the cells until ready for use in downstream experiments.
Therefore, the invention as a whole would have been prima facie obvious to a person of ordinary skill before the invention was filed.
Conclusion
No claims are allowed. No claims are free of the art.
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/Sean C. Barron/Primary Examiner, Art Unit 1653